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TRANSCRIPT
Formation and Cytotoxicity of Nanoparticles and Nanocubes
Using Gold and Silver Dan Banker, Skyler Dorrell, Prescott Ivey, Joe Scurti
Advisor: Dr. Tabbetha DobbinsDepartment of Physics and Astronomy, Rowan University
Agenda
Introduction
Preparation
Testing Methods
Cell Culture
Conclusion
Outlook
References
Introduction
Photothermal Therapy
Nanoparticles and Nanocubes of Noble Metals
Cytotoxicity
Analysis
Particles vs. Cubes, Gold vs. Silver
Huange X, Jain P, El-Sayed M, El-Sayed M. 2007. Plasmonic photothermal therapy (PPTT) using gold nanoparticles. Lasers Med Sci. [Springer-Verlag, London]
Preparation: Synthesis of Nanoparticles/Nanocubes
Preparation of Gold Nanoparticles
Boil a 50 mL aqueous solution of 1.0mM HAuCl4
Add a 5 mL aqueous solution of 38.8mM Na3C6H5O7
Boil until a dark red color becomes apparent
Preparation of Gold Nanocubes
Prepare a 10.05 mL aqueous growth solution with 2.5x10-4mM HAuCl4, 0.1M CTAB, and 0.1M ascorbic acid
Add 0.025 mL of gold nanoparticle seed solution*The production of silver nanoparticles and nanocubes is similar.Li Q, Cao Y. 2006. Preparation and Characterization of Gold Nanorods. [Hebei
University of Science and Technology, China]
Testing Methods: TEM
Used to approximate the size of individual particlesProduction of nanocubes, not nanorods
Gold nanocubesAuNP w/ double citrateBase AuNP
Zeta Potential
Surface charge of individual particles.Stability of the particles
Less charge = more agglomeration
CytotoxicityPositive Charge = Increased Cytotoxicity 90Plus Particle Size Analyzer
AuNC AuNP 2xC
AuNP
AgNC AgNP
UV-Vis.
Tests light absorption rangeConfirmation of gold and silver nanoparticle production (~420 nm for silver and
~520 nm for gold)Ideal laser wavelength for plasmon resonance
Genesis 10S UV-Vis. Spectrophotometer
AgNC AuNC
AgNP AuNP AuNP2XC
395518 514
526
Cell Culture Techniques (3T3)
Cells are kept in an incubator inside of the clean lab room
All work is done under the biosafety hood
To ensure cleanliness, everything is wiped down with 91% isopropyl alcohol before it enters the hood
Cell medium is made of DMEM, FBS, and Pen-Strep
Medium is heated to 37 degrees C
Cells are split into new plates with fresh medium twice a week (1:5 and 1:10 split)
Nanoparticles are centrifuged and added to medium for doping
Cytotoxicity by Visual Inspection (Control)
Day 1 Day 2 Day 3
Cytotoxicity by Visual Inspection (AuNP)
Day 2Day 1 Day 3
Cytotoxicity by Visual Inspection (AuNC)
Day 1 Day 3Day 2
Cytotoxicity by Visual Inspection (AgNP)
Day 2Day 1 Day 3
Cytotoxicity by Visual Inspection (AgNC)
Day 1 Day 3Day 2
Conclusion
Both types of nanoparticles hosted 3T3 cells well
Both types of nanocube solutions seemed to kill the cells, silver nanocubes not produced
Problem: Shape vs. Chemical Properties vs. Surface Charge?
Outlook
Protein Coated nanoparticles/nanocubes
High powered laser (more powerful than Raman)
Acknowledgements
Dr. Tabbetha Dobbins
Dr. Jeffrey Hettinger
Mr. Carl Lunk
References (1) Jana N, Gearheart L, Murphy C. 2001. Wet chemical synthesis of silver nanorods and nanowires of controllable aspect ratio [Cambridge, UK](2) Tong L, Wei Q, Wei A, Cheng J. 2009. Gold nanorods as contrast agents for biological imaging: optical properties, surface conjugation, and photothermal
effects. [West Lafayette, IN](3) Liu Y, Xu M, Chen Q, Guan G, Hu W, Zhao X, Qiao M, Hu H, Liang Y, Zhu H, et al. 2015. Gold nanorods/mesoporous silica-based nanocomposite as
theranostic agents for targeting near-infrared imaging and photothermal therapy induced with lazer. [Suzhou, China](4) Melancon M, Lu W, Yang Z, Zhang R, Cheng Z, Elliot A, Stafford J, Olsen T, Zhang J, Li C. 2008. In vitro and in vivo targeting of hollow gold nanoshells
directed at epidermal growth factor receptor for photothermal ablation therapy. [Santa Cruz, California](5) Mackey M, Ali M, Austin L, Near R, El-Sayed M. 2014. The most effective gold nanorod size for plasmonic photothermal therapy: Theory and in vitro
experiments. [Atlanta, Georgia](6) Khlebtsov B, Tuchin V, Khlebtsov N, Zharov V, Melnikov A. 2006. Optical amplification of photothermal therapy with gold nanoparticles and nanoclusters.
[Saratov, Russia](7) Huange X, Jain P, El-Sayed M, El-Sayed M. 2007. Plasmonic photothermal therapy (PPTT) using gold nanoparticles. Lasers Med Sci. [Springer-Verlag,
London](8) Li Q, Cao Y. 2006. Preparation and Characterization of Gold Nanorods. [Hebei University of Science and Technology, China](9) Barchiesi D, Kessentini S. 2010. Quantitative comparison of optimized nanorods, nanoshells, and hollow nanospheres for photothermal therapy. [Troyes
Cedex, France](10) Zhang X, Wu D, Shen X, Liu P, Yang N, Zhao B, Zhang H, Sun Y, Zhang L, Fan F. 2011. Size-dependent in vivo toxicity of PEG-coated gold nanoparticles
[Chinese Academy of Medical Sciences and Peking Union Medical College, Tianji](11) Agarwal A, Huang SW, O’Donnell M, Day KC, Day M, Kotov N, Ashkenazi S. 2007. Targeted gold nanorod contrast agent for prostate cancer detection by
photoacoustic imaging. [University of Michigan, Michigan](12) Liu Y, Ashton J, Moding E, Yuan H, Register J, Fales A, Choi J, Whitley M, Zhao X, Qi Yi, et al. 2015. A plasmonic gold nanostar theranostic probe for in
vivo tumor imaging and photothermal therapy. [Duke University, NC]
Questions?
Particle Size
Cluster size of nanoparticles and nanocubesAble to test the quality of the material
